H2S代谢基因3-MPST和SQOR在肺纤维化中的作用及其机制OA
Roles and mechanisms of the H2S metabolism genes 3-MPST and SQOR in pulmonary fibrosis
目的 探讨硫化氢(H2S)代谢基因3-巯基丙酮酸硫转移酶(3-MPST)和硫化物醌氧化还原酶(SQOR)在肺纤维化中的作用及其机制.方法 将40只SD大鼠随机分为对照组和模型组,分别在博来霉素处理后14和21 d取材,采用实时定量PCR和免疫组织化学法分析3-MPST和SQOR mRNA和蛋白在肺纤维化中表达的变化.将A549细胞随机分为对照组、TGF-β1组、TGF-β1+NaHS组、siRNA-3-MPST组、siRNA-3-MPST+TGF-β1组、siRNA-SQOR组和siRNA-SQOR+TGF-β1组.采用RNA干扰、实时定量PCR和Western blotting检测3-MPST、SQOR、α平滑肌肌动蛋白(α-SMA)、E-钙黏蛋白(E-cadherin)表达的变化,检测各组A549细胞中H2S和活性氧(ROS)水平的变化.结果 与对照组比较,模型组大鼠肺组织中3-MPST mRNA和蛋白表达水平降低,SQOR mRNA和蛋白表达水平升高(P<0.05).与对照组比较,TGF-β1组A549细胞中E-cadherin和3-MPST mRNA和蛋白表达水平以及H2S含量显著降低(P<0.05),α-SMA和SQOR mRNA和蛋白表达水平以及ROS水平显著升高(P<0.05).与TGF-β1组比较,TGF-β1+NaHS组上述变化减弱.与TGF-β1组比较,siRNA-3-MPST+TGF-β1组E-cadherin和SQOR mRNA和蛋白表达水平以及H2S含量显著降低(P<0.05),α-SMA mRNA和蛋白表达水平和ROS水平显著升高(P<0.05);siRNA-SQOR+TGF-β1组α-SMA、3-MPST mRNA和蛋白表达水平以及ROS水平显著降低(P<0.05),E-cadherin mRNA和蛋白表达水平和H2S含量显著升高(P<0.05).结论 3-MPST可能通过增加H2S含量、降低ROS水平发挥抑制上皮-间质转化和肺纤维化的作用,而SQOR可能通过减少H2S含量、提高ROS水平促进上皮-间质转化和肺纤维化.NaHS具有抑制上皮-间质转化和肺纤维化的作用.
Objective To explore the roles and underlying mechanisms of hydrogen sulfide(H2S)-metabolizing genes 3-mercaptopyruvate sulfurtransferase(3-MPST)and sulfide quinone oxidoreductase(SQOR)in pulmonary fibrosis.Methods Forty Sprague-Dawley rats were randomly divided into the control and bleomycin-induced pulmonary fibrosis model groups,and lung tissues were sampled at 14 and 21 days post-treatment.The expression levels of 3-MPST and SQOR in the rat lungs were assessed via immunohistochemistry and quantitative real-time PCR.In vitro,A549 cells were divided into the control,TGF-β1,TGF-β1+NaHS,siRNA-3-MPST,siRNA-3-MPST+TGF-β1,siRNA-SQOR,and siRNA-SQOR+TGF-β1 groups.The expression levels of 3-MPST,SQOR,α-SMA,and E-cadherin were assessed via Western blotting and quantitative real-time PCR.The content of H2S and the level of reactive oxygen species(ROS)were also determined.Results The expression level of 3-MPST was significantly downregulated,whereas the expression level of SQOR was upregulated in the fibrotic lungs of rats in the model group compared with those in the control group(P<0.05).Compared with A549 cells in the control group,the A549 cells in the TGF-β1 group displayed decreased levels of 3-MPST,E-cadherin,and H2S,and increased levels of SQOR,α-SMA,and ROS(P<0.05).These effects were attenuated by NaHS in the TGF-β1+NaHS group compared with those in the TGF-β1 group.Compared with the TGF-β1 group,the levels of E-cadherin,SQOR,and H2S in the siRNA-3-MPST+TGF-β1 group were significantly decreased(P<0.05),while the levels of α-SMA and ROS were significantly increased(P<0.05).In the siRNA-SQOR+TGF-β1 group,the levels ofα-SMA,3-MPST,and ROS were significantly decreased(P<0.05),while the level of E-cadherin and H2S were significantly increased(P<0.05).Conclusion 3-MPST appears to inhibit EMT and pulmonary fibrosis by increasing H2S content and reducing ROS level,whereas SQOR may promote fibrosis by decreasing H2S content and increasing ROS level.NaHS can also inhibit EMT and pulmonary fibrosis.
李聪;付明霞;王楠;柏雪莲;刘乃国
滨州医学院附属医院检验科,山东 滨州 256600||滨州医学院附属医院医学研究中心,山东 滨州 256600滨州市人民医院病理科,山东 滨州 256600滨州医学院附属医院医学研究中心,山东 滨州 256600滨州医学院附属医院医学研究中心,山东 滨州 256600滨州医学院附属医院检验科,山东 滨州 256600||滨州医学院附属医院医学研究中心,山东 滨州 256600
医药卫生
肺纤维化上皮-间质转化3-巯基丙酮酸硫转移酶硫化物醌氧化还原酶硫化氢代谢
pulmonary fibrosisepithelial-mesenchymal transition3-mercaptopyruvate sulfurtransferasesulfide quinone oxidoreduc-tasehydrogen sulfide metabolism
《中国医科大学学报》 2026 (2)
139-145,7
山东省自然科学基金(ZR2019MC019)
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